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. 2022 Feb 9;71(12):2539–2550. doi: 10.1136/gutjnl-2021-325150

Figure 7.

Figure 7

Follistatin-like protein 1 (FSTL1) deficiency attenuates inflammation and glycolysis in macrophages. Bone marrow-derived macrophages (BMDMs) treated with 100 ng/mL lipopolysaccharide (LPS) for 24 hours (A–G): (A–B) messenger RNA (mRNA) and protein expressions of tumour necrosis factor (TNF)-α, interleukin (IL)-1β and IL-10 were determined; (C) immunofluorescence staining of pyruvate kinase M2 (PKM2) and inducible nitric oxide synthase (iNOS); (D) protein expression of PKM2, p-PKM2, iNOS, p-STAT1 and p-STAT6; (E) mRNA expression of iNOS and Arg1 was measured; (F) immunofluorescence staining of p65 and DAPI; (G) protein expression of toll-like receptor (TLR)-4, p-nuclear factor kappa B (NF-κB) and p-Ikk; (H) lactate acid production was measured from BMDMs in response to LPS (100 ng/mL, 6 hours). (I) Glycolytic rate and compacities of BMDMs were measured by real-time recording of extracellular acidification rates (ECAR) after successively injection of glucose (Glc), oligomycin (O) and 2-DG. (J) FSTL1FL/FL and FSTL1M-KO BMDMs treated with LPS (100 ng/mL, 4 hours). These cells were washed twice and pretreated with 10 mM 2-DG for 1 hour, followed by treatment with or without LPS (100 ng/mL) for another 4 hours. mRNA expression of IL-1β and iNOS were determined. Data were presented as mean±SEM of at least three independent experiments; scale bars, 50 µm; *p<0.05, **p<0.01, ***p<0.001.